Detection of Mismatch Repair Deficiency and Microsatellite Instability in Colorectal Adenocarcinoma by Targeted Next-Generation Sequencing

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• 作者：Jonathan A. Nowak^&lowast ; ; Matthew B. Yurgelun^&dagger ; ; ^&Dagger ; ; Jacqueline L. Bruce^&lowast ; ; Vanesa Rojas-Rudilla^&lowast ; ; Dimity L. Hall^&lowast ; ; Priyanka Shivdasani^&lowast ; ; Elizabeth P. Garcia^&lowast ; ; Agoston T. Agoston^&lowast ; ; Amitabh Srivastava^&lowast ; ; Shuji Ogino^&lowast ; ; ^&Dagger ; ; ^§ ; ; Frank C. Kuo^&lowast ; ; Neal I. Lindeman^&lowast ; ; Fei Dong^&lowast ; ; ^{fdong1@partners.org}
• 刊名：The Journal of Molecular Diagnostics
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：19
• 期：1
• 页码：84-91
• 全文大小：607 K
• 卷排序：19

文摘

Mismatch repair protein deficiency (MMR-D) and high microsatellite instability (MSI-H) are features of Lynch syndrome–associated colorectal carcinomas and have implications in clinical management. We evaluate the ability of a targeted next-generation sequencing panel to detect MMR-D and MSI-H based on mutational phenotype. Using a criterion of >40 total mutations per megabase or >5 single-base insertion or deletion mutations in repeats per megabase, sequencing achieves 92% sensitivity and 100% specificity for MMR-D by immunohistochemistry in a training cohort of 149 colorectal carcinomas and 91% sensitivity and 98% specificity for MMR-D in a validation cohort of 94 additional colorectal carcinomas. False-negative samples are attributable to tumor heterogeneity, and next-generation sequencing results are concordant with analysis of microsatellite loci by PCR. In a subset of 95 carcinomas with microsatellite analysis, sequencing achieves 100% sensitivity and 99% specificity for MSI-H in the combined training and validation set. False-positive results for MMR-D and MSI-H are attributable to ultramutated cancers with POLE mutations, which are confirmed by direct sequencing of the POLE gene and are detected by mutational signature analysis. These findings provide a framework for a targeted tumor sequencing panel to accurately detect MMR-D and MSI-H in colorectal carcinomas.